kilr™ cytotoxicity assays and retroparticles
TRANSCRIPT
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KILR™ Cytotoxicity AssaysSpecifically Measure Target Cell Death in Co-cultures
22
Challenge For Developing Cancer Immunotherapies
• Specifically determine target cell death in co-cultures with immune effectors?
• How to develop assays with multiple target cells?
ADCC ADCP CAR-TT-cell Redirection
With Bi-Specific AbAdoptive T-Cell Therapy
33
Challenges with Existing Cytotoxicity Assays
• Dye-based assays are radioactive and/or leaky
• General cell death measures are not specific for target cells
• Assays may not reflect the true Mechanism of Action of the drug
• Low throughput of assays
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Introducing the KILR™ Cytotoxicity Assay
+
55
KILR™ Cytotoxicity Assay Highlights & Benefits
• Unparalleled Specificity• Signal only from dead target cells
• Easy to Use• Simple, add and read protocol with
chemiluminescent readout
• Exquisite Sensitivity• Detect as few as 75 dead cells/well
• Ultimate Flexibility• Run cytotoxicity assays from 30 minutes
to 72 hours
BenefitLow background and higher signal to noise ratios
66
KILR™ Cytotoxicity Assay Highlights & Benefits
• Unparalleled Specificity• Signal only from dead target cells
• Easy to Use• Simple, add and read protocol with
chemiluminescent readout
• Exquisite Sensitivity• Detect as few as 75 dead cells/well
• Ultimate Flexibility• Run cytotoxicity assays from 30 minutes
to 72 hours
Benefit
Rapid implementation in any lab & eliminate
radioactivity
77
KILR™ Cytotoxicity Assay Highlights & Benefits
• Unparalleled Specificity• Signal only from dead target cells
• Easy to Use• Simple, add and read protocol with
chemiluminescent readout
• Exquisite Sensitivity• Detect as few as 75 dead cells/well
• Ultimate Flexibility• Run cytotoxicity assays from 30 minutes
to 72 hours
ADCC assay with Cetuximab & PBMCs
Benefit
Low false negatives in a screen and better lead
optimization
88
KILR™ Cytotoxicity Assay Highlights & Benefits
• Unparalleled Specificity• Signal only from dead target cells
• Easy to Use• Simple, add and read protocol with
chemiluminescent readout
• Exquisite Sensitivity• Detect as few as 75 dead cells/well
• Ultimate Flexibility• Run cytotoxicity assays from 30 minutes
to 72 hours
Benefit
Develop assay time based on relevant drug biology
99
Broad Applications of KILR™ Assays
ADCC ADCP CAR-T T-cell RedirectionWith Bi-Specific Ab
Adoptive T-Cell Therapy
• Cell death endpoint
• Easy-to-use
• Reproducible for QC
• Cell death endpoint
• Plate-based assay
• Easy-to-use
• Target cell specific
• Target cell specific
• Run assay for 72 hrs
• Easy-to-use
• High throughput
• Reproducible for QC
• Target cell specific
• Run assay for 72 hrs
• Easy-to-use
• High throughput
• Reproducible for QC
• Target cell specific
• Run assay for 72 hrs
• High throughput
• Easy-to-use
• Reproducible for QC
Key Benefits
1010
KILR™ Applications Antibody Dependent Cellular Cytotoxicity
1 0 -1 2 1 0 -1 1 1 0 -1 0 1 0 -9 1 0 -8 1 0 -7 1 0 -6 1 0 -5
0
1 0
2 0
3 0
4 0
E rb B 2 O v e r-e x p re s s in g C e ll P o o ls
4 :1 N K : ta rg e t ce lls
T ra s tu z u m a b [g /m L ]%
ly
sis
S /BHillSlope EC50
SKBR3 2.113 2.654e-009
MCF7 1.330 8.779e-009
NCI N87 0.7026 1.110e-008
1 2 .2
6 .1
1 0 .8
1 0 -1 2 1 0 -1 1 1 0 -1 0 1 0 -9 1 0 -8 1 0 -7 1 0 -6 1 0 -5
0
3 0 0 0
6 0 0 0
9 0 0 0
U 2 O S -E G F R e P L
A n tib o d y [g /m L ]
RL
U
S /B
8 .6
HillSlope EC50
Cetuximab 1.021 4.696e-009
Rituximab ~ 19.81 ~ 5.119e-011 --
ADCC Assay Verified with Marketed Drugs
ADCC
1111 Data provided courtesy of Amgen
IC50 (ng/mL) Relative Potency R2
Ref. Std. 0.485 1 1
100% Ctrl. 0.519 0.961 0.999
60% 0.897 0.574 0.99980% 0.581 0.83 0.996
130% 0.421 1.21 0.997160% 0.324 1.64 0.998
KILR™ ApplicationsADCC for QC Lot Release Assays
Key Benefits
• True MOA of ADCC molecule
• Accurate
• Precise
• Reproducible
• Robust
1212
KILR™ ApplicationsAntibody Dependent Cellular Phagocytosis
ADCP with Primary Macrophages
Data provided courtesy of Janssen
1313
KILR™ ApplicationsBispecific Antibody for T-cell Redirection
Bi-specific AbPlate Target Cells in 384-well plate
Add non-activated primary T cells
Add antibody
Add KILR Detection reagent
Read signal
41 hours
1hour
Data provided courtesy of Janssen
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Summary & Available KILR™ Assays
• Highly-specific measure of target cell death in co-culture
• Easy to use with simple add and read protocol
• Highly sensitive detection of as few as 75 dead cells in a well
• Broad applications for immuno-oncology drug development
Product Part NumberKILR ARH-77 Cell Line 97-1001C017
KILR H322 Cell Pool 97-1003P020
KILR NCI-N87 Cell Pool 97-1004P021
KILR Raji Cell Pool 97-1012P026
KILR Ramos Cell Pool 97-1006P022
KILR SKBR3 Cell Pool 97-1002P018
KILR A549 Cell Pool 97-1005P015
KILR SKOV3 Cell Pool 97-1007P023
KILR NCI-H292 Cell Pool 97-1008P024
KILR Daudi Cell Pool 97-1009P025
Product Part NumberKILR Retroparticles for Suspension Cells (Neomycin) 97-0002
KILR Retroparticles for Adherent Cells (Neomycin) 97-0003
KILR Retroparticles for Adherent & Suspension Cells (Neomycin)
97-0004
Cell Lines Retrovirus To Make Your Own Cell Lines
